Real-time monitoring of protein conformational changes using a nano-mechanical sensor.

Détails

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Etat: Public
Version: de l'auteur
ID Serval
serval:BIB_FEFD9F5619A4
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Real-time monitoring of protein conformational changes using a nano-mechanical sensor.
Périodique
Plos One
Auteur(s)
Alonso-Sarduy L., De Los Rios P., Benedetti F., Vobornik D., Dietler G., Kasas S., Longo G.
ISSN
1932-6203 (Electronic)
ISSN-L
1932-6203
Statut éditorial
Publié
Date de publication
2014
Peer-reviewed
Oui
Volume
9
Numéro
7
Pages
e103674
Langue
anglais
Notes
Publication types: Journal Article Publication Status: epublish
Résumé
Proteins can switch between different conformations in response to stimuli, such as pH or temperature variations, or to the binding of ligands. Such plasticity and its kinetics can have a crucial functional role, and their characterization has taken center stage in protein research. As an example, Topoisomerases are particularly interesting enzymes capable of managing tangled and supercoiled double-stranded DNA, thus facilitating many physiological processes. In this work, we describe the use of a cantilever-based nanomotion sensor to characterize the dynamics of human topoisomerase II (Topo II) enzymes and their response to different kinds of ligands, such as ATP, which enhance the conformational dynamics. The sensitivity and time resolution of this sensor allow determining quantitatively the correlation between the ATP concentration and the rate of Topo II conformational changes. Furthermore, we show how to rationalize the experimental results in a comprehensive model that takes into account both the physics of the cantilever and the dynamics of the ATPase cycle of the enzyme, shedding light on the kinetics of the process. Finally, we study the effect of aclarubicin, an anticancer drug, demonstrating that it affects directly the Topo II molecule inhibiting its conformational changes. These results pave the way to a new way of studying the intrinsic dynamics of proteins and of protein complexes allowing new applications ranging from fundamental proteomics to drug discovery and development and possibly to clinical practice.
Pubmed
Web of science
Open Access
Oui
Création de la notice
05/09/2014 18:12
Dernière modification de la notice
20/08/2019 17:29
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